中国流行株HIV—1B亚型Gag重组鸡痘病毒构建及其免疫原性

在以鸡痘病毒(FPV)282E4株为基础构建的重组表达质粒pUTAL的ATI-P7.5复合启动子下游,插入编码中国流行株HIV-1 B亚型核心蛋白gag基因,构建了重组表达质粒pUTALG.重组质粒与FPV共转染CEF细胞,进行了同源重组.通过BUdR加压筛选,X-gal染色,获得重组鸡痘病毒vUTALG.Westernblot检测结果表明,重组病毒表达了Gag蛋白,裂解后其相对分子质量分别为55×103、48×103、24×l03和l 7×103.以重组病毒vUTALG免疫小鼠,与FPV对照比较,小鼠脾T淋巴细胞CD4+T细胞数和CD4+/CD8+值均有上升趋势,ConA、LPS诱导的脾细胞增殖能力增强,并能诱导CTL和HIV-1特异的血清抗体反应,说明该重组鸡痘病毒能提高小鼠细胞免疫和体液免疫水平.

Construction of Recombinant Fowlpox Virus of HIV-1 Capsid Protein Gag and Its Immunogenity

The gag gene of HIV-1 was inserted at downstream of the combined promoter in p UTAL,a expressing plasmid was constructed.By transfecting the plasmid into CEF cells pre-infected with FPV 2 82 E4 strain,a recombinantvirus v UTALG was obtained by selecting in the presence of BU- d R and subsequentblue plaque screening.Western blot showed that the recombinant virus expressed the Gag protein in the infected CEF cells.Mice were immunized with the recombinantvirus.The num- ber of CD4 cells from splenic lymphocytes increased and the proliferation response of splenocytes to Con A,LPSand CTL cytotoxicactivity elevated markedly with the HIV-1 -specific antibody responseto be induced.The results showed the recombinant virus could increase cellular and humoral immunity response in mice.